An antifungal protein was isolated from a culture of Bacillus subtilis strain B29. The isolation procedure comprised ion exchange chromatography on diethylaminoethyl （DEAE）-52 cellulose and gel filtration chromatogr...An antifungal protein was isolated from a culture of Bacillus subtilis strain B29. The isolation procedure comprised ion exchange chromatography on diethylaminoethyl （DEAE）-52 cellulose and gel filtration chromatography on Bio-Gel P-100. The protein was absorbed on DEAE-cellulose and Bio-Gel P-100. The purified antifungal fraction was designated as B29I, with a molecular mass of 42.3 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE）, pl value 5.69 by isoelectric focusing （IEF）-PAGE, and 97.81% purity by high performance liquid chromatography （HPLC）. B291 exhibited inhibitory activity on mycelial growth in Fusarium oxysporum, Rhizoctonia solani, Fusarium moniliforme, and Sclerotinia sclerotiorum. The 50% inhibitory concentrations （IC50） of its antifungal activity toward Fusarium oxysporum and Rhizoctonia solani were 45 and 112 μmol/L, respectively. B29I also demonstrated an inhibitory effect on conidial spore germination of Fusariurn oxysporum and suppression of germ-tube elongation, and induced distortion, tumescence, and rupture of a portion of the germinated spores.展开更多
The purpose of this study is to investigate the function of a novel potassium transporter gene (NrHAK1) isolated from Nicotiana rustica roots using yeast complement and real-time PCR technique. The complementary DNA (...The purpose of this study is to investigate the function of a novel potassium transporter gene (NrHAK1) isolated from Nicotiana rustica roots using yeast complement and real-time PCR technique. The complementary DNA (cDNA) of NrHAK1, 2 488 bp long, contains an open reading frame (ORF) of 2 334 bp encoding a protein of 777 amino acids (87.6 kDa) with 12 predicted transmembrane domains. The NrHAK1 protein shows a high sequence similarity to those of high-affinity potassium transporters in Mesembryanthemum, Phytolacca acinosa, Arabidopsis thaliana, and so on. We found that the NrHAK1 gene could complement the yeast-mutant defect in K+ uptake. Among several tissues surveyed, the expression level of NrHAK1 was most abundant in the root tip and was up-regulated when exposed to potassium starvation. Moreover, the transcript accumulation was significantly reduced by adding 5 mmol/L NH4+ to the solution. These results suggest that NrHAK1 plays an important role in potassium absorption in N. rustica.展开更多
The experiment was conducted to study the mutagenic effects of space environment on seedling growth in M1 generation and plant height and heading date in M2 generation in rice. Two types of lines of growth promotion ...The experiment was conducted to study the mutagenic effects of space environment on seedling growth in M1 generation and plant height and heading date in M2 generation in rice. Two types of lines of growth promotion （GP） and growth suppression （GS） were selected from the recombinant inbred lines （RILs） derived from a cross between a sensitive japonica Lemont and a nonsensitive indica Teqing to compare the mutation frequency and mutagenic efficiency for plant height and heading date in the M2 generation. Space environment resulted in 34.9% higher seedling height （SH） in the GP group than in the control, and 39.1% lower in the GS group than in the control, and there was no difference in seed fertility （SF） between the two groups. In M2, mutants of plant height and heading date can be induced by space treatment in both the two groups with lower mutation frequency and mutagenic efficiency in the GP group than in the GS group. There were no significant differences in the physiological damages in M1 between the two groups after γ-rays irradiation treatment. Mutation frequency and mutagenic efficiency of heading date in M2 were higher in the GS group than in the GP group, and the opposite was true for mutagenic efficiency of plant height although the mutation frequency varied between the two groups. The mutation frequencies of plant height and heading date induced by space environment were obviously lower than those by γ-rays irradiation, but the mutagenic efficiency was the opposite for most of the traits. For the GP and GS of seedlings induced by space environment, the GS had higher mutation frequency and mutagenic efficiency for plant height and heading date than the GP in M2.展开更多
Objective measurements are required for computer-aided sperm morphometric analysis （CASMA） machines to distinguish normal from abnormal sperm heads. The morphometric characteristics of spermatozoa in 72 samples of s...Objective measurements are required for computer-aided sperm morphometric analysis （CASMA） machines to distinguish normal from abnormal sperm heads. The morphometric characteristics of spermatozoa in 72 samples of semen and of spermatozoa from 72 other semen samples after swim-up were quantified by the semi-automated Integrated Sperm Analysis System （ISAS） computer-aided system, which measured the sperm head parameters length （L）, width （W）, area （A）, perimeter （P）, acrosomal area （Ac）, and the derived values L/W and P/A. For each man a homogeneous population of distributions characterized seminal spermatozoa （7 942 cells： median values L 4.4 μm, W 2.8μm A 9.8 μm^2, P 12.5 μm, Ac 47.5%, L/W 1.57, P/A 1.27）, and there was no significant difference in within- and among-individual variation. Different men could have spermatozoa of significantly different dimensions. Head dimensions for swim-up spermatozoa from different men （4 812 cells） were similar to those in semen, differing only by 2%-5%. The values of L, Wand L/W fell within the limits given by the World Health Organization （WHO）. Although these samples were not biologically matched, linear mixed-effects statistical analyses permitted valid comparison of the groups. A subpopulation of 404 spermatozoa considered to fit the stringent criteria of WHO ‘normal＇ seminal spermatozoa from both semen and swim-up were characterized by median values （and 95% confidence intervals） of L, 4.3 μm （3.8-4.9）, W, 2.9 μm （2.6-3.3）, A, 10.2μm^2 （8.5-12.2）, P, 12.4μm （11.3-13.9）, Ac, 49% （36-60）, L/W, 1.49 （1.32-1.67） and P/A, 1.22 （1.11-1.35）. These median values fall within the 95th centile confidence limits given by WHO, but the confidence intervals for L and W were larger. Although these differences in head dimensions among men and after swim-up could be detected by CASMA, the small differences make it unlikely that technicians would be able to distinguish them. The values could be used as default sperm head values for the CASMA machine used here.展开更多
Trichoderma spp. is a filamentous soil fungus known as an effective biocontrol agent of a range of important airborne and soilborne pathogens, it has universal distribution and economic importance. This article review...Trichoderma spp. is a filamentous soil fungus known as an effective biocontrol agent of a range of important airborne and soilborne pathogens, it has universal distribution and economic importance. This article reviewed the researches on biocontrol mechanism for plant diseases and application of Trichoderma spp., especially Trichoderma harzianum in recent years展开更多
Superoxide dismutases are metalloproteins which play a major role in defense against oxygen radicalmediated toxicity in aerobic organisms. Such proteins are important endogeneity cytoprotection factor involving defenc...Superoxide dismutases are metalloproteins which play a major role in defense against oxygen radicalmediated toxicity in aerobic organisms. Such proteins are important endogeneity cytoprotection factor involving defence. A 751-bp full-length cDNA sequence of an SOD gene was isolated from the Trichoderma harzianum. The full-length cDNA of the SOD gene consists of one 465-bp open reading frame nucleotide, which encodes a 15.7-kDa polypeptide consisting of 154 amino acid residues. Sequence analysis revealed that SOD gene has more than 72%-86% amino acid sequence homology with those of other fungi. The SOD gene was integrated into the genomic DNA of pYES2 by insertion into a single site for recombination, yielding the recombinant pYES2-SOD. SOD expressed by pYES2-SOD was induced by galactose. We test whether SOD could offer abiotic stress resistance when it was introduced into yeast ceils. A transgenic yeast harboring T. harzianum SOD was generated under the control of a constitutively expressed GAL promoter. The results indicated that SOD yeast transformants had significantly higher resistance to salt and drought stress.展开更多
A cDNA library was successfully constructed from Anip973,a human lung adenocarcinoma cell line with high metastatic potential.NIH3T3 cells were stably transfected using this cDNA library and screened for morphological...A cDNA library was successfully constructed from Anip973,a human lung adenocarcinoma cell line with high metastatic potential.NIH3T3 cells were stably transfected using this cDNA library and screened for morphological changes in a soft agar assay.Genomic DNA was isolated from putative clones and the integrated sequence was retrieved by PCR and sequencing.Three known genes,ribosomal protein L23,hypothetical protein FLJ22104,and serine protease inhibitor,kazal type 6 and a number of 5'-terminally truncated sequences were identified.Furthermore,cells transfected with ribosomal protein L23 was highly invasive compared with the empty vector as control(P〈0.02).These results indicate that the expression cloning of cDNA libraries in NIH3T3 cells and subsequent screening for loss of contact inhibition in soft agar is a viable tool for identifying tumor-related genes and ribosomal protein L23 gene plays a role in cell movement and metastasis.展开更多
A study was conducted on the molecular mechanism of small heat shock proteins （sHSPs） in Chaetomium globosum. Heat shock protein 22.4 （Hsp22.4） from C. globosum was cloned and expressed in Escherichia coli. BlastX...A study was conducted on the molecular mechanism of small heat shock proteins （sHSPs） in Chaetomium globosum. Heat shock protein 22.4 （Hsp22.4） from C. globosum was cloned and expressed in Escherichia coli. BlastX analysis revealed that the Hsp22.4 gene from C. globosum shared the highest identity in amino acid sequence with a Hsp gene from Neurospora crassa, and the identity between them was 65%. The C. globosum Hsp22.4 gene was inserted into the expressive vector of pGEX-4T-2 and the recombinant plasmid named pGEX-HSE E. coli BL21 transformed with pGEX-HSP plasmid was induced by IPTG, and the expressed proteins were analyzed with SDS-PAGE. A 50 kD protein was specially expressed in E. coli BL21, and the result was consistent with expectation, and showed that the Hsp22.4 gene had been expressed in E. coli. Our study has made a foundation for further studying the function ofsHSPs protein.展开更多
An atom gene was cloned from genomic DNA of Scleortinia sclerotiorum by inverse PCR. The evolutionary relationships of S. sclerotiorum and other fungi in atom gene were studied. Results showed that the atom gene from ...An atom gene was cloned from genomic DNA of Scleortinia sclerotiorum by inverse PCR. The evolutionary relationships of S. sclerotiorum and other fungi in atom gene were studied. Results showed that the atom gene from of S. sclerotiorum has a single open reading frame of 4 773 bp and does not include any introns. The derived amino acid sequence consists of 1 590 residues, and it is homologous to all fungal AROM proteins studied so far. The theoretical isoelectric point （pl） and molecular weight （Mw） is 6.5 and 172.66 kD, respectively GC percentage of the arom gene is 44.94. According to the results of searching from CDD and Prosite database, AROM protein of S. sclerotiorum contains five conserve domains： 3-dehydroquinate synthase domain, 3-dehydroquinate dehydratase （3-dehydroquinase） domain, shikimate 5-dehydrogenase domain, shikimate kinase domain, and -enolpyruvylshikimate-3-phosphate synthase （EPSP sythase） domain, and four motifs： two EPSP synthase signatures, dehydroquinase class I active site, shikimate kinase signature. According to the PIR Site Rule PIRSR000514-1, four functionally important amino acid residues are found by alignment. Putative TATA box and CAAT box locate separately in -23 and -77 loci in 5＇ un-translated region, and two loci found in downstream atom gene are likely polyadenylation signals. In addition, phylogeny of atom gene is analyzed.展开更多
Steam pretreatment was employed to disrupt Microalgal cells for lipids extraction.Effects of steam pretreatment on microstructure of microalgal cells were investigated through scanning electron microscopy(SEM)and tran...Steam pretreatment was employed to disrupt Microalgal cells for lipids extraction.Effects of steam pretreatment on microstructure of microalgal cells were investigated through scanning electron microscopy(SEM)and transmission electron microscopy(TEM).Effect of treatment on lipid extraction was also studied.Microalgal cell walls were distorted after steam pretreatment due to the hydrolysis of organic macromolecules contained in cell wall.Maximum curvature was increased from 1.88×10^(-6) m^(-1) to 1.43×10^(-7) m^(-1) after treatment with the steam at 130℃.The fractal dimension of microalgal cells increased from 1.25 to 1.30 after pretreatment for 15 min,and further increased to 1.47 when the pretreatment time was increased to 60 min.Increased steam pretreatment temperature and time enhanced the hydrolysis of organic macromolecules,and finally destroyed microalgal cell walls at pretreatment temperature of 130℃and pretreatment time of 60 min.Lipid extracted from wet microalgal was significantly increased(2.1-fold)after pretreatment.展开更多
For exploring the biological effect of space environment on different genotype of rice seeds, the cytological effects of M1 generation after space flight were studied. Twelve different genotypes of rice seeds which be...For exploring the biological effect of space environment on different genotype of rice seeds, the cytological effects of M1 generation after space flight were studied. Twelve different genotypes of rice seeds which belong to different climate ecotype （early, medium and late） of indica and japonica were onboard ＂Shenzhou 4＂ spaceship for 162 h. After recovered the total number of mitosis cells and chromosomal aberration were observed. In all the lines the mitotic index （MIs） of space flight are much higher than control, which indicates the stimulate effect of space environment. The cell rate of chromosomal aberration （CRCA） of space flight is also much higher than control, but varies from line to line. It indicates that biological effect of space environment on rice seed dependents not only on flight duration but also on rice genotype. The radiosensitivities of different lines were also discussed according to CRCAs.展开更多
The gene encoding the 20S proteasome subunit（PR29） was cloned from cDNA library of Trichoderma harzianum and expressed in Escherichia coli BL21 （D3） using a pET-28a expression system. The molecular weight of the p...The gene encoding the 20S proteasome subunit（PR29） was cloned from cDNA library of Trichoderma harzianum and expressed in Escherichia coli BL21 （D3） using a pET-28a expression system. The molecular weight of the protein was found to be approximately 29 kDa, as estimated by SDS-PAGE on gels. The target protein was insoluble when induced at 22℃ with 0.4 mmol/L IPTG, while dissoluble if induced at 37℃ with 0.8mmoL/L IPTG. The expressed product was purified through Ni-magnetic beads His Bind. The purity of the fusion protein reached above 80%. The entire eDNA sequence consisted of 1094 bp with 173 and 135 bp in 5＇ and 3＇ untranslated regions respectively. The gene encoding 261 amino acids has no signal peptide sequence. These results could provide a basis for validating the func-tions of PR29. It also provided a preliminary indication for further study of the mechanism and function of proteasome, and more information of proteasome mechanism in T.harzianum could be obtained.展开更多
It has been reported that Short staining provides additional morphological information on the motility of spermatozoa in semen,by distinguishing between red and blue flagella. With our routine methods （involving moun...It has been reported that Short staining provides additional morphological information on the motility of spermatozoa in semen,by distinguishing between red and blue flagella. With our routine methods （involving mounting slides） we were unable to confirm these observations. The presence of both red- and blue-coloured sperm tails in Short-stained semen smears was apparent,however,if slides were unmounted. Only a very weak association between blue flagellar staining and immotility was observed. Stating whether a mountant was used should be reported.展开更多
On-time mapping dynamics of crop area,yield,and production is important for global food security.Such information,however,is often not available.Here,we used satellite information,a spectral-phenology integration appr...On-time mapping dynamics of crop area,yield,and production is important for global food security.Such information,however,is often not available.Here,we used satellite information,a spectral-phenology integration approach for mapping crop area,and a machine learning model for predicting yield in the war-stricken Ukraine.We found that in Ukraine crop area and production declined in 2022 relative to 2017–2021 and 2021 for wintertriticeae crops,which was invaded before the cropping season in February of that year.At the same time,crop area and production for rapeseed increased in Ukraine,with yields consistently lower by 6.5%relative to 2021.The low precipitation and the Russian-Ukrainian conflict-related factors contributed to such yield variations by-1.3%and-0.9%for winter-triticeae crops and-4.2%and-0.5%for rapeseed in 2022.We demonstrate a robust framework for monitoring country-wide crop production dynamics in near real-time,serving as an early-foodsecurity-warning system.展开更多
One mutant line eto with salt tolerance was screened from a T-DNA insertion mutant collection of Arabidopsis thaliana. In addition to a reduced rate of seed germination, NaCl and ABA also inhibited the growth and the ...One mutant line eto with salt tolerance was screened from a T-DNA insertion mutant collection of Arabidopsis thaliana. In addition to a reduced rate of seed germination, NaCl and ABA also inhibited the growth and the greening of cotyledons of wild-type seedlings, but not the eto mutant. TAIL-PCR analysis showed that T-DNA tag insertion in the eto was located at nucleotide 27,502 in BAC F3M18, upstream （at position -487 relative to the translation initiation codon） of gene At lg77740 （encoding a putative phosphatidylinositol-4-phosphate 5-kinase, AtPIP5K2）. This inserted mutation cosegregated closely with the eto phenotype, Another analysis not only indicated that AtPIP5K2 transcript is expressed predominantly in roots and rosette leaves, but also showed the T-DNA insertion resulted higher accumulation of the AtPIP5K2 in eto mutant plants and did not influenced the expression of the upstream At lg77730 gene. This change may play an essential role in the tolerance of eto mutant plant to the osmotic stress.展开更多
Imaging technologies based on chemiluminescence(CL)and bioluminescence(BL)have seen a tremendous growth in the past decade due to their extensive contributions to biochemical analysis and biomedical science.CL and BL ...Imaging technologies based on chemiluminescence(CL)and bioluminescence(BL)have seen a tremendous growth in the past decade due to their extensive contributions to biochemical analysis and biomedical science.CL and BL imaging technologies have many advantages over commonly used imaging techniques(such as fluorescence and electrochemical systems).CL and BL proceed without an external light source,thus avoiding photobleaching,high interference of background signal,and autofluorescence.Furthermore,CL and BL analytical techniques are characterized by their low detection limit,high selectivity,short assay time,and simple instrumentation.Recently,CL-based imaging technology has been applied successfully for the determination and in vivo imaging of several significant analytes.Meanwhile,an innovative BL imaging technology has been established for the noninvasive real-time tracking of different biomolecules relying on the interaction between luciferase and its substrate.In the current review,we discuss the recent applications of CL and BL imaging approaches over the past five years.Finally,we also discuss the current state of progress and the prospects for CL and BL imaging systems.展开更多
Background A20, also known as tumor necrosis factor α induced protein 3 （TNFaip3）, is a cytoplasmic zinc finger protein that inhibits nuclear factor kappa-B （NF-κB） activity and prevents tumor necrosis factor （...Background A20, also known as tumor necrosis factor α induced protein 3 （TNFaip3）, is a cytoplasmic zinc finger protein that inhibits nuclear factor kappa-B （NF-κB） activity and prevents tumor necrosis factor （TNF）-mediated programmed cell death. NF-κB is a transcription factor that regulates expression of genes involved in cell proliferation, cell survival and anti-apoptosis. Several studies have implicated that the NF-κB signal pathway is associated with angiogenesis and clinico-pathological process of adenoid cystic carcinoma （ACC） of the salivary glands.Methods The ability of overexpression of A2.0 to influence the biological behavior and invasion of ACC cells was examined. The cells were stably transfected with full-length A20 cDNA. Stable gene transfer was verified by realtime-polymerase chain reaction （PCR） and Western blot analysis. The change of cell biological behavior was examined by methyl thiazolyl tetrazolium （MTT） and NF-κB luciferase reporter assay and the invasion of the cells was examined by a Matrigel invasion chamber.Results pEGPFN3-A2.0 gene was stably transferred into ACC-2 cells and overexpressed. When cells were treated with TNFα, the NF-κB activity of ACC-2-A20 cells could be down-regulated about 46.32% in contrast to ACC-2-GFP cells （P〈0.05）. A20 potently inhibited growth of A20 transfectant ACC-2-A20 compared with control vector transfected groups and the ACC-2 empty control group （P〈0.05）. The ACC-2-A20 cells showed significantly reduced ability to invade through Matdgei-coated filters compared to ACC-2-GFP and ACC-2 cells. The inhibition rate was up to 71.05% （P〈0.05）. Conclusions A2.0 gene transfer is associated with decreased tumor invasion, in part via the down-regulation of NF-κB expression, providing evidence for a potential application of A20 in designing a treatment modality for salivary gland cancers such as ACC.展开更多
基金supported by the Hi-Tech Research and Development Pro-gram (863) of China (No. 2003AA241140)the Natural Science Foundation of Heilongjiang Province, China (No. C200522)
文摘An antifungal protein was isolated from a culture of Bacillus subtilis strain B29. The isolation procedure comprised ion exchange chromatography on diethylaminoethyl （DEAE）-52 cellulose and gel filtration chromatography on Bio-Gel P-100. The protein was absorbed on DEAE-cellulose and Bio-Gel P-100. The purified antifungal fraction was designated as B29I, with a molecular mass of 42.3 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE）, pl value 5.69 by isoelectric focusing （IEF）-PAGE, and 97.81% purity by high performance liquid chromatography （HPLC）. B291 exhibited inhibitory activity on mycelial growth in Fusarium oxysporum, Rhizoctonia solani, Fusarium moniliforme, and Sclerotinia sclerotiorum. The 50% inhibitory concentrations （IC50） of its antifungal activity toward Fusarium oxysporum and Rhizoctonia solani were 45 and 112 μmol/L, respectively. B29I also demonstrated an inhibitory effect on conidial spore germination of Fusariurn oxysporum and suppression of germ-tube elongation, and induced distortion, tumescence, and rupture of a portion of the germinated spores.
基金(No. 110200101008) supported by the State Tobacco Mo-nopoly Administration, China
文摘The purpose of this study is to investigate the function of a novel potassium transporter gene (NrHAK1) isolated from Nicotiana rustica roots using yeast complement and real-time PCR technique. The complementary DNA (cDNA) of NrHAK1, 2 488 bp long, contains an open reading frame (ORF) of 2 334 bp encoding a protein of 777 amino acids (87.6 kDa) with 12 predicted transmembrane domains. The NrHAK1 protein shows a high sequence similarity to those of high-affinity potassium transporters in Mesembryanthemum, Phytolacca acinosa, Arabidopsis thaliana, and so on. We found that the NrHAK1 gene could complement the yeast-mutant defect in K+ uptake. Among several tissues surveyed, the expression level of NrHAK1 was most abundant in the root tip and was up-regulated when exposed to potassium starvation. Moreover, the transcript accumulation was significantly reduced by adding 5 mmol/L NH4+ to the solution. These results suggest that NrHAK1 plays an important role in potassium absorption in N. rustica.
文摘The experiment was conducted to study the mutagenic effects of space environment on seedling growth in M1 generation and plant height and heading date in M2 generation in rice. Two types of lines of growth promotion （GP） and growth suppression （GS） were selected from the recombinant inbred lines （RILs） derived from a cross between a sensitive japonica Lemont and a nonsensitive indica Teqing to compare the mutation frequency and mutagenic efficiency for plant height and heading date in the M2 generation. Space environment resulted in 34.9% higher seedling height （SH） in the GP group than in the control, and 39.1% lower in the GS group than in the control, and there was no difference in seed fertility （SF） between the two groups. In M2, mutants of plant height and heading date can be induced by space treatment in both the two groups with lower mutation frequency and mutagenic efficiency in the GP group than in the GS group. There were no significant differences in the physiological damages in M1 between the two groups after γ-rays irradiation treatment. Mutation frequency and mutagenic efficiency of heading date in M2 were higher in the GS group than in the GP group, and the opposite was true for mutagenic efficiency of plant height although the mutation frequency varied between the two groups. The mutation frequencies of plant height and heading date induced by space environment were obviously lower than those by γ-rays irradiation, but the mutagenic efficiency was the opposite for most of the traits. For the GP and GS of seedlings induced by space environment, the GS had higher mutation frequency and mutagenic efficiency for plant height and heading date than the GP in M2.
文摘Objective measurements are required for computer-aided sperm morphometric analysis （CASMA） machines to distinguish normal from abnormal sperm heads. The morphometric characteristics of spermatozoa in 72 samples of semen and of spermatozoa from 72 other semen samples after swim-up were quantified by the semi-automated Integrated Sperm Analysis System （ISAS） computer-aided system, which measured the sperm head parameters length （L）, width （W）, area （A）, perimeter （P）, acrosomal area （Ac）, and the derived values L/W and P/A. For each man a homogeneous population of distributions characterized seminal spermatozoa （7 942 cells： median values L 4.4 μm, W 2.8μm A 9.8 μm^2, P 12.5 μm, Ac 47.5%, L/W 1.57, P/A 1.27）, and there was no significant difference in within- and among-individual variation. Different men could have spermatozoa of significantly different dimensions. Head dimensions for swim-up spermatozoa from different men （4 812 cells） were similar to those in semen, differing only by 2%-5%. The values of L, Wand L/W fell within the limits given by the World Health Organization （WHO）. Although these samples were not biologically matched, linear mixed-effects statistical analyses permitted valid comparison of the groups. A subpopulation of 404 spermatozoa considered to fit the stringent criteria of WHO ‘normal＇ seminal spermatozoa from both semen and swim-up were characterized by median values （and 95% confidence intervals） of L, 4.3 μm （3.8-4.9）, W, 2.9 μm （2.6-3.3）, A, 10.2μm^2 （8.5-12.2）, P, 12.4μm （11.3-13.9）, Ac, 49% （36-60）, L/W, 1.49 （1.32-1.67） and P/A, 1.22 （1.11-1.35）. These median values fall within the 95th centile confidence limits given by WHO, but the confidence intervals for L and W were larger. Although these differences in head dimensions among men and after swim-up could be detected by CASMA, the small differences make it unlikely that technicians would be able to distinguish them. The values could be used as default sperm head values for the CASMA machine used here.
基金Supported by Chinese National Programs for High Technology Research and Development(2003AA241140)
文摘Trichoderma spp. is a filamentous soil fungus known as an effective biocontrol agent of a range of important airborne and soilborne pathogens, it has universal distribution and economic importance. This article reviewed the researches on biocontrol mechanism for plant diseases and application of Trichoderma spp., especially Trichoderma harzianum in recent years
基金the National High Technology Research and Development Programme of China(No.2003AA241140)
文摘Superoxide dismutases are metalloproteins which play a major role in defense against oxygen radicalmediated toxicity in aerobic organisms. Such proteins are important endogeneity cytoprotection factor involving defence. A 751-bp full-length cDNA sequence of an SOD gene was isolated from the Trichoderma harzianum. The full-length cDNA of the SOD gene consists of one 465-bp open reading frame nucleotide, which encodes a 15.7-kDa polypeptide consisting of 154 amino acid residues. Sequence analysis revealed that SOD gene has more than 72%-86% amino acid sequence homology with those of other fungi. The SOD gene was integrated into the genomic DNA of pYES2 by insertion into a single site for recombination, yielding the recombinant pYES2-SOD. SOD expressed by pYES2-SOD was induced by galactose. We test whether SOD could offer abiotic stress resistance when it was introduced into yeast ceils. A transgenic yeast harboring T. harzianum SOD was generated under the control of a constitutively expressed GAL promoter. The results indicated that SOD yeast transformants had significantly higher resistance to salt and drought stress.
基金This work was supported by Returning Scholars Fund of Heilongjiang Province(No.LC04C02)the Department of Education Overseas Researcher Fund of Heilongjiang Province(No.1054HZ013).
文摘A cDNA library was successfully constructed from Anip973,a human lung adenocarcinoma cell line with high metastatic potential.NIH3T3 cells were stably transfected using this cDNA library and screened for morphological changes in a soft agar assay.Genomic DNA was isolated from putative clones and the integrated sequence was retrieved by PCR and sequencing.Three known genes,ribosomal protein L23,hypothetical protein FLJ22104,and serine protease inhibitor,kazal type 6 and a number of 5'-terminally truncated sequences were identified.Furthermore,cells transfected with ribosomal protein L23 was highly invasive compared with the empty vector as control(P〈0.02).These results indicate that the expression cloning of cDNA libraries in NIH3T3 cells and subsequent screening for loss of contact inhibition in soft agar is a viable tool for identifying tumor-related genes and ribosomal protein L23 gene plays a role in cell movement and metastasis.
文摘A study was conducted on the molecular mechanism of small heat shock proteins （sHSPs） in Chaetomium globosum. Heat shock protein 22.4 （Hsp22.4） from C. globosum was cloned and expressed in Escherichia coli. BlastX analysis revealed that the Hsp22.4 gene from C. globosum shared the highest identity in amino acid sequence with a Hsp gene from Neurospora crassa, and the identity between them was 65%. The C. globosum Hsp22.4 gene was inserted into the expressive vector of pGEX-4T-2 and the recombinant plasmid named pGEX-HSE E. coli BL21 transformed with pGEX-HSP plasmid was induced by IPTG, and the expressed proteins were analyzed with SDS-PAGE. A 50 kD protein was specially expressed in E. coli BL21, and the result was consistent with expectation, and showed that the Hsp22.4 gene had been expressed in E. coli. Our study has made a foundation for further studying the function ofsHSPs protein.
文摘An atom gene was cloned from genomic DNA of Scleortinia sclerotiorum by inverse PCR. The evolutionary relationships of S. sclerotiorum and other fungi in atom gene were studied. Results showed that the atom gene from of S. sclerotiorum has a single open reading frame of 4 773 bp and does not include any introns. The derived amino acid sequence consists of 1 590 residues, and it is homologous to all fungal AROM proteins studied so far. The theoretical isoelectric point （pl） and molecular weight （Mw） is 6.5 and 172.66 kD, respectively GC percentage of the arom gene is 44.94. According to the results of searching from CDD and Prosite database, AROM protein of S. sclerotiorum contains five conserve domains： 3-dehydroquinate synthase domain, 3-dehydroquinate dehydratase （3-dehydroquinase） domain, shikimate 5-dehydrogenase domain, shikimate kinase domain, and -enolpyruvylshikimate-3-phosphate synthase （EPSP sythase） domain, and four motifs： two EPSP synthase signatures, dehydroquinase class I active site, shikimate kinase signature. According to the PIR Site Rule PIRSR000514-1, four functionally important amino acid residues are found by alignment. Putative TATA box and CAAT box locate separately in -23 and -77 loci in 5＇ un-translated region, and two loci found in downstream atom gene are likely polyadenylation signals. In addition, phylogeny of atom gene is analyzed.
基金supported by the National Key Research and Development Program-China(2017YFE0122800)Shandong Provincial Natural Science Foundation(ZR2019MC060)+1 种基金Key Research and Development Program of Jining City(2018ZDGH024)a Project of Shandong Province Higher Educational Science and Technology Program(J17KA095)。
文摘Steam pretreatment was employed to disrupt Microalgal cells for lipids extraction.Effects of steam pretreatment on microstructure of microalgal cells were investigated through scanning electron microscopy(SEM)and transmission electron microscopy(TEM).Effect of treatment on lipid extraction was also studied.Microalgal cell walls were distorted after steam pretreatment due to the hydrolysis of organic macromolecules contained in cell wall.Maximum curvature was increased from 1.88×10^(-6) m^(-1) to 1.43×10^(-7) m^(-1) after treatment with the steam at 130℃.The fractal dimension of microalgal cells increased from 1.25 to 1.30 after pretreatment for 15 min,and further increased to 1.47 when the pretreatment time was increased to 60 min.Increased steam pretreatment temperature and time enhanced the hydrolysis of organic macromolecules,and finally destroyed microalgal cell walls at pretreatment temperature of 130℃and pretreatment time of 60 min.Lipid extracted from wet microalgal was significantly increased(2.1-fold)after pretreatment.
基金Sponsored by the National"863"Program Project (2004AA231071) the Ministerial Level Foundation
文摘For exploring the biological effect of space environment on different genotype of rice seeds, the cytological effects of M1 generation after space flight were studied. Twelve different genotypes of rice seeds which belong to different climate ecotype （early, medium and late） of indica and japonica were onboard ＂Shenzhou 4＂ spaceship for 162 h. After recovered the total number of mitosis cells and chromosomal aberration were observed. In all the lines the mitotic index （MIs） of space flight are much higher than control, which indicates the stimulate effect of space environment. The cell rate of chromosomal aberration （CRCA） of space flight is also much higher than control, but varies from line to line. It indicates that biological effect of space environment on rice seed dependents not only on flight duration but also on rice genotype. The radiosensitivities of different lines were also discussed according to CRCAs.
文摘The gene encoding the 20S proteasome subunit（PR29） was cloned from cDNA library of Trichoderma harzianum and expressed in Escherichia coli BL21 （D3） using a pET-28a expression system. The molecular weight of the protein was found to be approximately 29 kDa, as estimated by SDS-PAGE on gels. The target protein was insoluble when induced at 22℃ with 0.4 mmol/L IPTG, while dissoluble if induced at 37℃ with 0.8mmoL/L IPTG. The expressed product was purified through Ni-magnetic beads His Bind. The purity of the fusion protein reached above 80%. The entire eDNA sequence consisted of 1094 bp with 173 and 135 bp in 5＇ and 3＇ untranslated regions respectively. The gene encoding 261 amino acids has no signal peptide sequence. These results could provide a basis for validating the func-tions of PR29. It also provided a preliminary indication for further study of the mechanism and function of proteasome, and more information of proteasome mechanism in T.harzianum could be obtained.
文摘It has been reported that Short staining provides additional morphological information on the motility of spermatozoa in semen,by distinguishing between red and blue flagella. With our routine methods （involving mounting slides） we were unable to confirm these observations. The presence of both red- and blue-coloured sperm tails in Short-stained semen smears was apparent,however,if slides were unmounted. Only a very weak association between blue flagellar staining and immotility was observed. Stating whether a mountant was used should be reported.
基金supported by the National Natural Science Foundation of China(Grant No.42061144003).
文摘On-time mapping dynamics of crop area,yield,and production is important for global food security.Such information,however,is often not available.Here,we used satellite information,a spectral-phenology integration approach for mapping crop area,and a machine learning model for predicting yield in the war-stricken Ukraine.We found that in Ukraine crop area and production declined in 2022 relative to 2017–2021 and 2021 for wintertriticeae crops,which was invaded before the cropping season in February of that year.At the same time,crop area and production for rapeseed increased in Ukraine,with yields consistently lower by 6.5%relative to 2021.The low precipitation and the Russian-Ukrainian conflict-related factors contributed to such yield variations by-1.3%and-0.9%for winter-triticeae crops and-4.2%and-0.5%for rapeseed in 2022.We demonstrate a robust framework for monitoring country-wide crop production dynamics in near real-time,serving as an early-foodsecurity-warning system.
基金Supported by the National Program for Transgenic Plants from China （ GN. J99-A-001 ） and National Natural Science Foundation of China （ GN. 30221120261 ）.
文摘One mutant line eto with salt tolerance was screened from a T-DNA insertion mutant collection of Arabidopsis thaliana. In addition to a reduced rate of seed germination, NaCl and ABA also inhibited the growth and the greening of cotyledons of wild-type seedlings, but not the eto mutant. TAIL-PCR analysis showed that T-DNA tag insertion in the eto was located at nucleotide 27,502 in BAC F3M18, upstream （at position -487 relative to the translation initiation codon） of gene At lg77740 （encoding a putative phosphatidylinositol-4-phosphate 5-kinase, AtPIP5K2）. This inserted mutation cosegregated closely with the eto phenotype, Another analysis not only indicated that AtPIP5K2 transcript is expressed predominantly in roots and rosette leaves, but also showed the T-DNA insertion resulted higher accumulation of the AtPIP5K2 in eto mutant plants and did not influenced the expression of the upstream At lg77730 gene. This change may play an essential role in the tolerance of eto mutant plant to the osmotic stress.
基金support by National Natural Science Foundation of China(22174136,22004116,and 21874126)Department of Science and Technology of Jilin Province(L202234)+3 种基金Chinese Academy of Sciences(CAS),Iran National Science Foundation(CAS-VPST Silk Road Science Fund 2021 Nos.GJHZ202125 and INSF99008701)the Ministry of Education and Science of Ukraine(M/109-2019,M/97-2020,0122U001533)National Research Foundation of Ukraine(2020.02/0390)the Ministry of Science and Technology of the People’s Republic of China(CU03-02).
文摘Imaging technologies based on chemiluminescence(CL)and bioluminescence(BL)have seen a tremendous growth in the past decade due to their extensive contributions to biochemical analysis and biomedical science.CL and BL imaging technologies have many advantages over commonly used imaging techniques(such as fluorescence and electrochemical systems).CL and BL proceed without an external light source,thus avoiding photobleaching,high interference of background signal,and autofluorescence.Furthermore,CL and BL analytical techniques are characterized by their low detection limit,high selectivity,short assay time,and simple instrumentation.Recently,CL-based imaging technology has been applied successfully for the determination and in vivo imaging of several significant analytes.Meanwhile,an innovative BL imaging technology has been established for the noninvasive real-time tracking of different biomolecules relying on the interaction between luciferase and its substrate.In the current review,we discuss the recent applications of CL and BL imaging approaches over the past five years.Finally,we also discuss the current state of progress and the prospects for CL and BL imaging systems.
基金Chinese National Natural and Scientific Committee(No.30330580)
文摘Background A20, also known as tumor necrosis factor α induced protein 3 （TNFaip3）, is a cytoplasmic zinc finger protein that inhibits nuclear factor kappa-B （NF-κB） activity and prevents tumor necrosis factor （TNF）-mediated programmed cell death. NF-κB is a transcription factor that regulates expression of genes involved in cell proliferation, cell survival and anti-apoptosis. Several studies have implicated that the NF-κB signal pathway is associated with angiogenesis and clinico-pathological process of adenoid cystic carcinoma （ACC） of the salivary glands.Methods The ability of overexpression of A2.0 to influence the biological behavior and invasion of ACC cells was examined. The cells were stably transfected with full-length A20 cDNA. Stable gene transfer was verified by realtime-polymerase chain reaction （PCR） and Western blot analysis. The change of cell biological behavior was examined by methyl thiazolyl tetrazolium （MTT） and NF-κB luciferase reporter assay and the invasion of the cells was examined by a Matrigel invasion chamber.Results pEGPFN3-A2.0 gene was stably transferred into ACC-2 cells and overexpressed. When cells were treated with TNFα, the NF-κB activity of ACC-2-A20 cells could be down-regulated about 46.32% in contrast to ACC-2-GFP cells （P〈0.05）. A20 potently inhibited growth of A20 transfectant ACC-2-A20 compared with control vector transfected groups and the ACC-2 empty control group （P〈0.05）. The ACC-2-A20 cells showed significantly reduced ability to invade through Matdgei-coated filters compared to ACC-2-GFP and ACC-2 cells. The inhibition rate was up to 71.05% （P〈0.05）. Conclusions A2.0 gene transfer is associated with decreased tumor invasion, in part via the down-regulation of NF-κB expression, providing evidence for a potential application of A20 in designing a treatment modality for salivary gland cancers such as ACC.
